摘要
为了研究抗Ⅰ型糖尿病融合蛋白质基因HSP65-6×P277在植物中的表达,构建了含有融合基因HSP65-6×P277的植物表达载体pCAMBIA2301-HSP65-6×P277,通过农杆菌介导法将重组载体转入烟草,利用卡那霉素作为植物筛选标记,获得含有抗Ⅰ型糖尿病基因的16株转基因烟草植株。通过PCR、半定量RT-PCR及SDS-PAGE蛋白质电泳等方法检测,初步验证在抗性烟草植株中融合基因HSP65-6×P277在mRNA和蛋白质水平上进行了表达。
With the purpose of studying the expression of recombinant gene HSP65-6xP277 in plant, the plant consti- tutive expression vector pCAMBIA2301-HSP65-6xP277 containing fusion gene HSP65-6xP277 was constructed and trans- formed into tobacco plants via Agrobacterium tumefaciens. Sixteen resistant plants containing anti-type I diabetes gene were obtained by screening with kanamysin. The resistant plants were analyzed by PCR, RT-PCR and SDS-PAGE. The results showed that the fusion gene HSP65-6xP277 was expressed in the resistant tobacco plants on DNA, mRNA and protein levels.
出处
《江苏农业学报》
CSCD
北大核心
2012年第3期477-481,共5页
Jiangsu Journal of Agricultural Sciences
基金
国家转基因专项(2011ZX08005-001、2012ZX0801-003)
江苏省“333”工程科研项目(005035310922)
关键词
Ⅰ型糖尿病
P277肽
烟草
转化
植物疫苗
type I diabetes
peptide P277
tobacco
transformation
plant vaccinum
